Sufficient quantities of enzymes and proteins (carbonic anhydrase, phosphopyruvate hydratase, serine dehydratase, phospho-transacetylase, alcoholic dehydrogenase, aldolase) and their apo derivatives will be prepared and characterized to allow appropriate thermodynamic and C13 NMR studies to proceed. Log K, delta H degrees, and delta S degrees values will be determined at 25 degrees by a calorimetric titration procedure for the reaction of Hplus and OHminus with the enzymes and proteins and their apo derivatives mentioned previously. These data will provide information concerning the number and type of metal binding groups and reaction stoichiometry. These determinations will be repeated at 37 degrees for selected systems making possible the calculation of delta Cp degrees values. In addition interactions of metal free bovine and human serum albumins with selected transition metal ions will be studied calorimetrically at 37 degrees. These delta H degrees data will be combined with those previously determined at 25 degrees to calculate delta Cp degrees values for these systems. Calorimetric titration data will also be collected for the reaction at 25 degrees of Hplus and OHminus with carbonic anhydrase where Zn has been replaced by Cd2 plus , Cu2 plus, Ni2 plus, Fe2 plus, and Mn2 plus. Carbon-13 NMR data will be obtained for selected metal-protein systems selected from those outlined above. These data will be used in conjunction with the calorimetric data to identify specific metal binding sites.